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首页> 外文期刊>Journal of Virology >Biological and biochemical studies of cells transformed by simian virus 40 temperature-sensitive gene A mutants and A mutant revertants.
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Biological and biochemical studies of cells transformed by simian virus 40 temperature-sensitive gene A mutants and A mutant revertants.

机译：Simian病毒40温度敏感基因变异的细胞生物和生化研究突变体和突变体还原剂。

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摘要

The growth properties of hamster cells transformed by wild-type Simian virus 40 (SV40), by early SV40 temperature-sensitive mutants of the A complementation group, and by spontaneous revertants of these mutants were studied. All of the tsA mutant-transformed cells were temperature sensitive in their ability to form clones in soft agar and on monolayers of normal cells except for CHLA-30L1, which was not temperature sensitive in the latter property. All cells transformed by stable revertants of well-characterized tsA mutants possessed certain growth properties in common with wild-type-transformed cells at both temperatures. Virus rescued from tsA transformants including CHLA30L1 was temperature sensitive for viral DNA replication, whereas that rescued from revertant and wild-type transformants was not thermolabile in this regard. T antigen present in crude extracts of tsA-transformed cells including CHLA30L1, grown at 33 degreeC, was temperature sensitive by in vitro immunoassay, whereas that from wild-type-transformed cells was relatively stable. T antigen from revertant transformants was more stable than the tsA protein. Partially purified T antigen from revertant-transformed cells was nearly as stable as wild-type antigen in its ability to bind DNA after heating at 44 degrees C, whereas T antigen from tsA30 mutant-transformed cells was relatively thermolabile. These results further indicate that T antigen is a product of the SV40 A gene. Significantly more T antigen was found in extracts of CHLA30L1 grown to high density at the nonpermissive temperature than in any other tsA-transformed cell similarly grown. This is consistent with the suggestion that the amount of T antigen synthesized in CHLA30L1 is large enoughto allow partial expression of the transformed phenotype at the restrictive temperature. Alternatively, the increase in T antigen concentration may be secondary to one or more genetic alterations that independently affect the transformed phenotype of these cells.
机译：通过互补组的早期SV40温度敏感突变体和通过这些突变体的自发恢复剂，研究了野生型猿猴病毒40（SV40）转化的仓鼠细胞的生长特性。所有TSA突变体转化的细胞在它们在软琼脂中形成克隆的能力和除CHLA-30L1之外的正常细胞中的单层敏感的能力敏感，其在后一种物质中没有温度敏感。通过稳定的TSA突变体的稳定恢复剂转化的所有细胞具有在两个温度下的野生型转化的细胞中具有某些生长性质。从包括CHLA30L1的TSA转化体救出的病毒对病毒DNA复制温度敏感，而从回避和野生型转化体中救出的病毒在这方面拯救了野生型转化体并不热压。在包括CHLA30L1的TSA转化细胞的粗提取物中存在的T抗原，在33℃下生长，通过体外免疫测定温度敏感，而来自野生型转化的细胞相对稳定。来自回复剂转化体的抗原比TSA蛋白更稳定。部分纯化的来自转化体转化的细胞的T抗原几乎与其在44℃加热后结合DNA的野生型抗原的稳定性，而来自TSA30突变体转化细胞的T抗原相对热。这些结果进一步表明T抗原是SV40 A基因的产物。在非智能温度下生长至高密度的CHLA30L1的提取物中发现了更多的T抗原大于同样地生长的任何其他TSA转化的细胞。这与建议一致的是，在CHLA30L1中合成的T抗原的量是大的，允许在限制温度下允许转化的表型的部分表达。或者，T抗原浓度的增加可以是一种或多种遗传改变，其独立地影响这些细胞的转化表型。

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